Molecular Characterization of the D Surface Protein Gene Subfamily in Paramecium tetraurelia

ABSTRACT. When Paramecium tetraurelia expresses the D serotype, detectable by serum tests, high molecular mRNA could be isolated, which corresponds to the molecular mass of the D surface protein. Using this D specific mRNA as a probe for screenings in different genomic libraries a subfamily of five very similar genes was found, named α-51D, _γ1-51D, _γ2-51D, δ-51D and ε-51D. Each of them is about 8-kb long, they show regions of identity to each other, and there is no evidence that any are defective genes or pseudogenes. Up to now serotype D is the only known serotype showing this phenomenon. Another novel feature is that two of the D isogenes are closely linked. The sequence for the entire coding region of the α-51D gene has been determined, as well as the upstream and downstream noncoding regions. Its deduced amino acid sequence shows the same characteristic cysteine periodicity displayed by all other immobilization antigen (i-ag) genes from Paramecium. However, in contrast to most other such genes, tandem repeats are missing from the 7599-bp long coding region of the α-51D gene. When the sequences of the type 51D genes are compared to each other, the similarity is very high and extends to coding as well as to noncoding regions. Similarity within noncoding regions is usually only observed for allelic i-ag genes. We conclude that the type D genes constitute a family of isogenes that are nonallelic. They contain slightly different consensus sequences with possible functions as regulatory regions.     

Homozygosity at a class II MHC locus depresses female reproductive ability in European brown hares

The link between adaptive genetic variation, individual fitness and wildlife population dynamics is fundamental to the study of ecology and evolutionary biology. In this study, a Bayesian modelling approach was employed to examine whether individual variability at two major histocompatibility complex (MHC) class II loci (DQA and DRB) and eight neutral microsatellite loci explained variation in female reproductive success for wild populations of European brown hare (Lepus europaeus). We examined two aspects of reproduction: the ability to reproduce (sterility) and the number of offspring produced (fecundity). Samples were collected from eastern Austria, experiencing a sub‐continental climatic regime, and from Belgium with a more Atlantic‐influenced climate. As expected, reproductive success (both sterility and fecundity) was significantly influenced by age regardless of sampling locality. For Belgium, there was also a significant effect of DQA heterozygosity in determining whether females were able to reproduce (95% highest posterior density interval of the regression parameter [−3.64, −0.52]), but no corresponding effect was found for Austria. In neither region was reproduction significantly associated with heterozygosity at the DRB locus. DQA heterozygotes from both regions also showed a clear tendency, but not significantly so, to produce a larger number of offspring. Predictive simulations showed that, in Belgium, sub‐populations of homozygotes will have higher rates of sterile individuals and lower average offspring numbers than heterozygotes. No similar effect is predicted for Austria. The mechanism for the spatial MHC effect is likely to be connected to mate choice for increased heterozygosity or to the linkage of certain MHC alleles with lethal recessives at other loci.     

Spatial and temporal variation in the biomass and nutrient status of epilithic algae in Lake Erken, Sweden

1. The aim of this study was to estimate patchiness in biomass and in the internal nutrient status of benthic algae on hard substrata (epilithon) in Lake Erken, Sweden, over different levels of distance, depth and time. Knowledge of the sources and scale of patchiness should enable more precise estimation of epilithic biomass and nutrient status for the entire lake. We focused on the horizontal scale, about which little is known. 2. We sampled epilithon by SCUBA diving and used a hierarchical sampling design with different horizontal scales (cm, dm, 10 m, km) which were nested in two temporal scales (within and between seasons). We also compared two successive years and three sampling depths (0, 1 and 4 m). Biomass was measured as particulate carbon and chlorophyll a (Chl a) and internal nutrient status as carbon : nitrogen : phosphorus (C : N : P) ratios and as specific alkaline phosphatase activity (APA). 3. Horizontal variation accounted for 60–80 and 7–70% of the total variation in biomass and in nutrient status, respectively, at all depths and during both years. Both small and large scales accounted for significant variation. We also found variation with time and depth. Biomass increased in autumn after a summer minimum, and the within‐season variation was very high. The lowest biomass was found at 0 m depth. Both N and P limitation occurred, being higher in 1996 than in 1997 and decreased with depth. 4. As a consequence, any sampling design must address variation with distance, depth and time when estimating biomass or nutrient limitation of benthic algae for an entire lake. Based on this analysis, we calculated an optimal sampling design for detecting change in the epilithic biomass of Lake Erken between different sampling days. It is important to repeat the sampling as often as possible, but also the large scales (10 m and km) and the dm scale should be replicated. Using our calculations as an example, and after a pilot study, an optimal sampling design can be computed for various objectives and for any lake. 5. Short‐term impact of the wind, light and nutrient limitation, and grazing, might be important in regulating the biomass and nutrient status of epilithic algae in Lake Erken. Patchiness in the nutrient status of algae was not coupled to the patchiness of biomass, indicating that internal nutrients and biomass were regulated by different factors.     

New Methods for Cultivating,Harvesting, and Purifying Mass Cultures of the Hypotrich Ciliate Euplotes aediculatus1

A new method is described for mass cultivation of Euplotes aediculatus, a hypotrich ciliate containing omikron-symbionts. The ciliate cultures, continuously aerated in Erlenmeyer flasks (5000 ml) with 4500 ml medium, yield densities of 2300 cells/ml which are four to five times higher than cell densities of cultures grown in unaerated Fernbach flasks. Harvesting such cultures involves the application of 25-μm mesh sieves. Cells so concentrated can be purified by using columns or special chambers in which Euplotes migrates towards the cathodes in an electric field (field strength 7 V/cm).     

Contribution of biofilm‐dwelling consumers to pelagic–benthic coupling in a large river

1. Over the course of this 17‐month study, we assessed the potential loss of plankton (bacteria, algae, heterotrophic flagellates) to consumers (ciliates and rotifers) within mature biofilms established on natural substrata exposed to the main current of the River Rhine (Germany). Once a month, in flow cells in a bypass system to the River Rhine, we measured the clearance rates of the biofilm‐associated consumers on the different groups within the natural plankton. 2. Ciliates were the most dominant consumers, among which planktivorous groups, particularly peritrichs and (in spring and summer) heterotrichs dominated. Consumer biomass varied with season, with the highest density occurring directly after the appearance of the phytoplankton spring peak. 3. Clearance rates on plankton ranged from 96 to 565 L m^?2 d^?1 for bacteria and 66–749 L m^?2 d^?1 for algae, with a preference for algae in summer and for bacteria in winter. This pattern coincided with seasonal changes in the structures of the grazer communities. The consumers (both ciliates and rotifers with total standing stocks ranging between 19 and 572 mg C m^?2) imported a substantial amount of organic matter (between 15 and 137 mg C m^?2 d^?1) into the biofilm. 4. These results highlight the potential importance of consumers in the biofilm as a trophic link between the plankton and the benthos, a function that has hitherto mostly been attributed to filter‐feeding bivalves. In contrast to bivalves, the biofilm‐dwelling consumers show a more dynamic response towards the plankton density and composition. Such dynamic components need to be considered when estimating total plankton consumption by the benthos.     

Selectivity and competitive interactions between two benthic invertebrate grazers (Asellus aquaticus and Potamopyrgus antipodarum): an experimental study using 13C- and 15N-labelled diatoms

1. Tracer experiments with two diatoms labelled with ¹³C (Nitzschia palea) and ¹⁵N (Fragilaria crotonensis), were conducted to investigate feeding selectivity and interspecific competition between the grazers Asellus aquaticus (Isopoda, Crustacea) and Potamopyrgus antipodarum (Hydrobiidae, Gastropoda). Conventional methods, such as cell counts and estimated biovolume, were used first to detect feeding preferences within the different grazer treatments. 2. The results revealed a significant decline in algal biovolume in all grazer treatments and no indications of active selectivity were observed. In contrast to conventional methods, measurements based on isotope signatures showed strong differences in tracer uptake, thus indicating different degrees of assimilation and digestion by the two grazers. 3. The selectivity index Q, which provides information on the uptake ratio of ¹³C to ¹⁵N, showed a significant time effect for both grazer species and a significant difference between single‐ and mixed‐grazer treatments for P. antipodarum. Thus, this technique enabled the direct quantification of the uptake by grazers and, therefore, served as an ideal tool for the detection of passive selectivity. 4. Our results indicate a shift in feeding preferences related to between‐species competition and a potential divergence of trophic niches when species coexist.     

Variation of distal genitalia in the simultaneously hermaphroditic land snail Arianta arbustorum (Pulmonata,Stylommatophora) caused by sexual selection?

The most likely explanation for genitalic extravagance may be sexual selection acting either before, during, or after copulation. In promiscuous species, the intensity of postulating sexual selection may be a function of population density. In this study we examined the variability of the distal genitalia involved in spcrmatophore production, reception and manipulation of 113 adult individuals of the simultaneously hermaphroditic land snail Arianta arbustorum (L.) from six natural populations in the Eastern Alps (Gesause, Austria). We investigated the hypothesis that these genitalia increase in relation to shell size with increasing population density (range: 0.9 to 39.8 individuals/m²) and expected a higher variance of the genitalia compared to shell dimensions due to sexual selection. Genitalic size was unexpectedly inversely related to population density, probably due to increased inhibitor)^- effects of snail mucus. Patterns of variation of female and male characters did not differ. Coefficients of variation of the genitalia were significantly higher than those of the shell dimensions as predicted. This was due to a higher dispersion around the regression lines rather than higher allometric values. However, the influence of sexual selection on genitalic size and variance cannot be unambiguously determined. We discuss different scenarios emphasizing the importance of sexual selection to differing degrees and conclude that sexual selection has probably played only a minor role.     

Low algal carbon content and its effect on the C : P stoichiometry of periphyton

1. We examined the contribution of algal cells to periphytic organic carbon and assessed the effects of variable biomass composition on the carbon : phosphorus (C : P) ratio of periphyton. We compiled more than 5000 published and unpublished observations of periphytic carbon : chlorophyll a (C : Chl) ratios, an index of algal prevalence, from a variety of substrata collected from lake and low‐salinity coastal habitats. In addition, we converted estimates of algal biovolume into algal C to obtain an independent measure of cellular algal carbon in periphyton. This information was used in a model relating periphyton C : P ratio to algal cellular carbon, the algal C : P ratio, and the C : P ratio of non‐algal organic matter in periphyton. 2. The mean C : Chl ratio of periphyton (405) was relatively high with values in >25% of the samples exceeding 500. On average, 8.4% of total periphyton C was accounted for by C in algal cells. Only 15% of samples were found to have more than 15% periphyton C in cellular algal carbon. Our model showed a nonlinear relationship between periphytic C : P ratios and the C : P ratio of algal cells in the periphyton when non‐algal organic matter was present. However, even at relatively low cellular algal C (<10% of total C), algal C : P ratios can strongly affect the C : P ratio of periphyton as a whole (i.e. algal cells plus other organic matter). 3. The high C : Chl ratios and the low biovolume‐derived algal C of periphyton samples in our data set indicate that algal cells are typically a minor component of organic carbon in periphyton, However, this minor contribution would not preclude algal cellular stoichiometry from notably influencing periphyton C : P ratios.